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Biosci Biotechnol Biochem ; 88(8): 892-899, 2024 Jul 22.
Article in English | MEDLINE | ID: mdl-38830810

ABSTRACT

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has infected many people around the world; fast and accurate detection of the virus can help control the spread of the virus. Reverse transcription-polymerase chain reaction (RT-PCR) is the gold standard method for SARS-CoV-2 detection. In this study, we improved the RT-PCR by proposing a novel method using dual double-quenched fluorescence probes. We used the improved probes to detect the plasmid DNA and RNA reference materials of SARS-CoV-2, respectively. The results show that, the background fluorescence intensity reduced by 50%, the fluorescence increment increased to 2.8 folds, and the Ct value significantly reduced by 3 or more, indicating that the detection sensitivity increased at least 8 times. In addition, we demonstrated that the improved probes have well performance in detecting SARS-CoV-2, with the minimum concentration of 6.2 copies/µL. This study will help biological companies develop better products for SARS-CoV-2 and other clinical pathogen infection.


Subject(s)
COVID-19 , Fluorescent Dyes , RNA, Viral , SARS-CoV-2 , SARS-CoV-2/isolation & purification , SARS-CoV-2/genetics , Fluorescent Dyes/chemistry , Humans , COVID-19/diagnosis , COVID-19/virology , RNA, Viral/analysis , RNA, Viral/genetics , Sensitivity and Specificity , Reverse Transcriptase Polymerase Chain Reaction/methods , COVID-19 Nucleic Acid Testing/methods , Limit of Detection
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